Team:Hawaii/Notebook/2008-07-29

From 2008.igem.org

(Difference between revisions)
(Wetlab work)
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:<strong>Grace</strong>
:<strong>Grace</strong>
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:* Purified RE digests on EtBr stained 3% agarose gel; extracted desired bands
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:* Purified RE digests on EtBr stained 3% agarose gel
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:* Ligated pRL1383a with ccdB, nir with B0030, and GFP and GFPf with B0024
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::* Extracted bands for B0030 and B0024
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::* Other lanes FAILED!
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===Transformation of constructs in DB3.1===
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:<strong>Grace</strong>
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===Direct Band Extraction Method===
===Direct Band Extraction Method===

Revision as of 04:04, 30 July 2008

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Things we did today

Wetlab work

Gel purification of ccdB PCR product

Grace

RE digests

Grace
  • Digested pRL1383a (from last night) with BamHI
  • Digested B0030 and B0024 (from last night) with EcoRI
  • Sequentially digested gel purified ccdB with HindIII and BamHI

Construction of BioBricks

Grace
  • Purified RE digests on EtBr stained 3% agarose gel
  • Extracted bands for B0030 and B0024
  • Other lanes FAILED!

Direct Band Extraction Method

  • make two comb gels
  • run test UPA band along with ladder
  • attempt to pipette out DNA as it runs into the second well
  • no more gel cutting???

PCR amplification of pRL1383a parts

Margaret
  • Made a reaction buffer containing {38.5ul, 20ul accusure, 10ul dNTPs, & 5ul 5XBuffer}. This should be good for 20 50ul reactions.
  • Amplified with Accusure: aadA ([http://partsregistry.org/Part:BBa_J23012 BBa_J23012]), aadaA (pRL1383a), rep region, oriV
  • Amplified with Green Taq: rep region

Media

  • started to make LB amp100 plates, but autoclave room is closed. Remember to do it tomorrow.

Drylab Work

Name of Task

name of person/people who performed the task
  • Summary of task and what was done. Link to experiment for detailed notes if necessary.
  • e.g. read through papers, worked on proposal, etc.


Discussion

Quote of the Day

LOL! http://www.youtube.com/watch?v=J0s0Y3-BCaw